Abstract
BackgroundChikungunya (CHIKV) virus is an important mosquito-borne virus causing outbreaks of acute febrile illness with arthropathy. The detection of specific antibodies against CHIKV is used for diagnosis after the acute viremic phase of the disease. However, a major challenge for serologic diagnosis of CHIKV and other alphaviruses is the cross-reactivity of antibodies to common antigens among these viruses. In the present study, we have developed an enzyme-linked immunosorbend assay using a recombinant envelope protein 2 of CHIKV produced in Escherichia coli system, as a capture antigen.ResultsHigh titers (1600 to 12,800) of anti-CHIKV antibodies were detected in human sera analyzed by the CHIKV assay, suggesting it may detect low levels of the antibodies presence. On the other side, cross-reactivity was not observed in mouse hyperimmune sera to Mayaro virus and other alphaviruses analyzed by the CHIKV immunosorbend assay, suggesting it is a CHIKV-specific test. Fifty-nine human serum samples of CHIKV infection suspected cases were tested for immunoglobulin G (IgG) and M (IgM) antibodies detection using the CHIKV immunosorbend assay. A total of 44% (26/59) of samples were positive for IgG to CHIKV, determining 89.66% sensitivity and 100% specificity when the assay is compared to a CHIKV-specific neutralization assay. In addition, 40.6% (24/59) of samples were positive for IgM, determining 92.48% sensitivity and 79.04% specificity by a Bayesian method in the absence of a gold standard. Moreover, CHIKV immunosorbend assay showed similar sensibilities to a commercial immunochromatography assay (Lumiquick, USA) for CHIKV IgG and IgM detection.ConclusionIn short, we have developed a rapid, simple, specific and sensitive CHIKV immunosorbend assay for IgG and IgM detection and our results showed potential applicability on the diagnosis of infections by this virus.
Highlights
Chikungunya (CHIKV) virus is an important mosquito-borne virus causing outbreaks of acute febrile illness with arthropathy
Standardization of recombinant envelope protein 2 (rE2)-Chikungunya virus (CHIKV) Enzyme-linked immunosorbend assay (ELISA) A suitable amount of 44 kDa rE2 of CHIKV was produced in E. coli cells, and its purity was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot (Fig. 1a and b)
Cross-reactivity of other mouse hyperimmune sera was not observed for alphaviruses, such as Aura virus (AURV), Eastern equine encephalitis virus (EEEV), Mayaro virus (MAYV), Mucambo virus (MUCV), and Western equine encephalitis virus (WEEV) in the rE2-CHIKV ELISA (Fig. 2)
Summary
Chikungunya (CHIKV) virus is an important mosquito-borne virus causing outbreaks of acute febrile illness with arthropathy. The detection of specific antibodies against CHIKV is used for diagnosis after the acute viremic phase of the disease. A major challenge for serologic diagnosis of CHIKV and other alphaviruses is the cross-reactivity of antibodies to common antigens among these viruses. Chikungunya virus (CHIKV) is included in the Semliki Forest group of the Alphavirus genus (Togaviridae family). CHIKV is a mosquito-borne virus that causes human disease mainly characterized by acute onset fever and prominent arthropathy. CHIKV infection can cause persistent arthropathy for weeks to years, leading to incapacitation of patients and substantial economic loss [4]. CHIKV was first isolated from an acute febrile human case in 1953 during a Dengue epidemic in Liteho city, Tanzania [5].
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